Claims:

1. A method for preparing
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide as a maleate salt comprising the
step of: mixing
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide and maleic acid in a water-alcohol
solution at an elevated temperature.

2. The method of claim 1, further comprising the steps of: cooling said
solution to precipitate said maleate salt and filtering said solution
after cooling to obtain crystalline
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide maleate.

3. The method of claim 1 or 2, wherein said elevated temperature is from
about 40.degree. C. to about 60.degree. C.

4. The method of claim 2 or 3, wherein said cooling continues until the
solution reaches a temperature of about 37.degree. C. or less.

5. The method of claim 4, wherein said cooling continues until the
solution reaches a temperature of about 30.degree. C. or less.

6. The method of any one of claims 1-5, wherein said water-alcohol
solution comprises from about 5% to about 20% by volume water and from
about 80% to about 95% by volume alcohol.

7. The method of any one of claims 1-6, wherein said alcohol is
n-propanol.

8. The method of claim 7, wherein said water-alcohol solution comprises
about 10% by volume water and about 90% by volume n-propanol.

9. The method of any one of claims 1-8, wherein said solution further
comprises at least one compound selected from the group consisting of:
##STR00017## ##STR00018## ##STR00019##

10. The method of any one of claims 2-9, wherein said crystalline
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide maleate comprises crystals in
anhydrous form, monohydrate form, or combinations thereof.

11. A pharmaceutical composition for the inhibition of HER-2 kinase
activity comprising, a therapeutically-effective amount of
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide maleate and a pharmaceutically
acceptable carrier.

12. The pharmaceutical composition according to claim 11 wherein said
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide maleate is in a form selected from
the group consisting of: an anhydrous form, a monohydrate form, and
combinations thereof.

13. The pharmaceutical composition according to claim 11 or 12 further
comprising at least one compound selected from the group consisting of:
##STR00020## ##STR00021## ##STR00022##

14. A compound selected from the group consisting of: ##STR00023##
##STR00024## ##STR00025##

15. A method for preventing, treating, or inhibiting cancer comprising,
administering to a subject a therapeutically-effective amount of
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide maleate.

17. A method of preparing
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide maleate monohydrate comprising the
steps selected from the group consisting of: a. dissolving anhydrous
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide maleate with an organic solvent,
adding water, and filtering; b. dissolving anhydrous
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide maleate with an organic solvent,
adding a solution of a second organic solvent and water, and filtering;
c. dissolving anhydrous
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide maleate with an organic solvent
containing water, and filtering; and d. reslurrying anhydrous
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide maleate with an organic solvent
containing water for a period of days, and filtering.

18. The method of claim 17, wherein the steps are dissolving anhydrous
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide maleate with an organic solvent,
adding water and filtering.

19. The method of claim 18, wherein the organic solvent is
dimethylsulfoxide.

20. The method of claim 18 or 19 further comprising heating the solution
to between about 45.degree. C. to about 60.degree. C.

21. The method of claim 17, wherein the steps are dissolving anhydrous
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide maleate with an organic solvent,
adding a solution of a second organic solvent and water and filtering.

22. The method of claim 21, wherein the organic solvent is
dimethylsulfoxide, the second organic solvent is selected from the group
consisting of tetrahydrofuran, isopropanol, n-propanol, acetone, ethanol,
methanol, and acetonitrile, and the amount of water present is about 7.5%
to about 15% by weight.

23. The method of claim 17, wherein the steps are dissolving anhydrous
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide maleate with an organic solvent
containing water and filtering.

24. The method of claim 23, wherein the organic solvent is selected from
the group consisting of tetrahydrofuran, isopropanol, n-propanol,
acetone, ethanol, methanol, and acetonitrile, and the amount of water
present is about 7.5% to about 20% by weight.

25. The method of claim 24, wherein the organic solvent is n-propanol and
the amount of water present is about 10% to about 15% by weight.

26. The method of claim 17, wherein the steps are: reslurrying anhydrous
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide maleate with an organic solvent
containing water for a period of days, and filtering.

27. The method of claim 26, wherein the organic solvent is
tetrahydrofuran and the period of days is about 10-20.

28. A method of preparing anhydrous
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide maleate comprising drying under
vacuum monohydrate
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide maleate at a temperature greater
than 30.degree. C. for about 12 to about 48 hours.

32. An isolated crystalline form according to claim 31 having
substantially the X-ray diffraction pattern as shown in FIG. 6.

33. An isolated crystalline form of anhydrous
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide maleate, characterized by an onset
temperature of about 196.degree. C. and ending temperature about
204.degree. C., as measured by differential scanning calorimetry.

34. An isolated crystalline form of
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide maleate monohydrate, having a water
content of about 2.5 to 2.7%, by weight.

35. An isolated crystalline form of partially hydrated
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide maleate, having a water content of
about 1.5% to about 2.3%, by weight.

Description:

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit under 35 U.S.C. §119(e) to
co-pending U.S. Provisional Application Ser. No. 61/124,796, filed Oct.
17, 2007, which is hereby incorporated by reference in its entirety.

FIELD OF THE INVENTION

[0002] This invention is directed to maleate salts of
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide, crystalline forms thereof, methods
of preparing the salts, associated compounds, pharmaceutical compositions
containing the maleate salt, and methods for their use. Maleate salts of
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide are useful in the treatment of
cancer.

BACKGROUND OF THE INVENTION

[0003] Compounds derived from 3-cyanoquinoline have been shown to have
anti-tumor activity, which may make them useful as chemotherapeutic
agents in treating various cancers, including but not limited to,
pancreatic cancer, melanoma, lymphatic cancer, parotid tumors. Barrett's
esophagus, esophageal carcinomas, head and neck tumors, ovarian cancer,
breast cancer, epidermoid tumors, cancers of major organs, such as
kidney, bladder, larynx, stomach, and lung, colonic polyps and colorectal
cancer and prostate cancer. Examples of compounds derived from
3-cyanoquinoline are disclosed and shown to possess anti-tumor activity
in U.S. Pat. Nos. 6,002,008; 6,432,979; and 6,288.082. One limitation of
certain 3-cyanoquinoline compounds is that they are not water soluble in
a free base form.

[0004] The crystalline form of a particular drug as a salt, a hydrate
and/or any polymorph thereof is often one important determinant of the
drug's ease of preparation, stability, water solubility, storage
stability, ease of formulation and in-vivo pharmacology. It is possible
that one crystalline form is preferable over another where certain
aspects such as ease of preparation, stability, water solubility and/or
superior pharmacokinetics are deemed to be critical. Crystalline forms of
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide salts that possess a higher degree
of water solubility than the free base but are stable fulfill an unmet
need for stable, crystalline, water-soluble forms of substituted
3-cyanoquinoline compounds that selectively inhibit kinase activity,
which in turn inhibit cell proliferation and tumorigenesis.

SUMMARY OF THE INVENTION

[0005] The present invention provides crystalline forms of
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide maleate, which have been isolated
and characterized as: an anhydrous form, a monohydrate form, and a
mixture of the anhydrous and the monohydrate forms (referred to as a
partial hydrate form). The invention is also directed to methods for
using this maleate salt and the crystalline forms thereof, and
pharmaceutical formulations containing them.

[0006] The invention provides an isolated crystalline form of anhydrous
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide maleate (Form I), characterized by
differential scanning calorimetry (DSC), as exhibiting an onset
temperature in the range of about 196-204° C., at which melting
and decomposition occur.

[0008] The invention provides an isolated crystalline form of
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide maleate monohydrate (Form II),
exhibiting water loss at about 50° C. and characterized by a water
content of about 2.5 to 2.7% by weight, based on the weight of the
compound as a monohydrate.

[0009] The invention also provides an isolated crystalline form of
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide maleate monohydrate (Form II),
wherein the maleate salt is characterized by XRD peaks at the following
angles (±0.20°) of 20 in its X-ray diffraction pattern: 6.53,
8.43, 10.16, 12.19, 12.47, 13.01, 15.17, 16.76, 17.95, 19.86, 21.11,
21.88, 23.22, 23.78, 25.69, 26.17, 27.06, 27.58, 28.26, 28.73, and 29.77.
In a separate embodiment, the isolated crystalline form of
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide maleate monohydrate exhibits an
X-ray diffraction pattern wherein all of the X-ray diffraction peaks are
at about the 2θ angles disclosed above.

[0010] The invention also provides an isolated crystalline form of
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide maleate monohydrate (Form II),
characterized by DSC, as exhibiting an onset temperature in the range of
196-204° C., at which melting and decomposition occur, especially
at a transition temperature of about 203.8° C.

[0011] The invention provides an isolated crystalline form of a partially
hydrated (E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-etho-
xy-6-quinolinyl}-4-(dimethylamino)-2-butenamide maleate (Form III),
characterized by a water content of about 0.8 to about 2.4% by weight,
including about 1.5% to about 2.3% by weight, based on the weight of the
compound.

[0012] The present invention provides a method of preparing the maleate
salt by mixing
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide (the free base) with maleic acid
and dissolving the mixture in a water-alcohol solution at an elevated
temperature. The resulting solution is cooled and the cooled solution
contains (E)-N{-4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-etho-
xy-6-quinolinyl}-4-(dimethylamino)-2-butenamide maleate.

[0013] The invention also provides a method of preparing
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide maleate in the form of a
crystalline monohydrate (Form II) comprising the steps of: mixing
anhydrous (E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-eth-
oxy-6-quinolinyl}-4-(dimethylamino)-2-butenamide maleate (Form I) with an
organic solvent and an amount of water and filtering crystalline
monohydrate that precipitates from the mixture.

[0014] The invention also provides a method of preparing
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide maleate in the form of a
crystalline monohydrate (Form II) comprising the steps of: mixing
anhydrous (E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-eth-
oxy-6-quinolinyl}-4-(dimethylamino)-2-butenamide maleate (Form I) with an
organic solvent; adding a solution comprising an amount of water in an
organic solvent; and filtering crystalline monohydrate that precipitates
from the mixture.

[0015] The invention also provides a method of preparing
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide maleate in the form of a
crystalline monohydrate (Form II) comprising the steps of: mixing
anhydrous (E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-eth-
oxy-6-quinolinyl}-4-(dimethylamino)-2-butenamide maleate (Form I) with an
organic solvent and an amount of water and filtering crystalline
monohydrate that precipitates from the mixture.

[0016] The invention also provides a method of preparing
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide maleate in the form of a
crystalline monohydrate (Form II) comprising the steps of: mixing
anhydrous (E)-N-{-4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-et-
hoxy-6-quinolinyl}-4-(dimethylamino)-2-butenamide maleate (Form I) with an
organic solvent comprising an amount of water and filtering crystalline
monohydrate that precipitates from the mixture.

[0017] The invention also provides a method of preparing
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide maleate in the form of a
crystalline monohydrate (Form II) comprising the steps of: mixing
anhydrous (E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-eth-
oxy-6-quinolinyl}-4-(dimethylamino)-2-butenamide maleate (Form I) with an
organic solvent comprising an amount of water over a period of days and
filtering crystalline monohydrate that precipitates from the mixture.

[0018] The invention also provides a method of preparing
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide maleate in anhydrous form (Form I)
comprising the step of: drying under vacuum
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide maleate as a monohydrate (Form II)
at a temperature greater than 30° C. for about 12 to about 48
hours.

[0019] The invention also provides a pharmaceutical formulation
comprising:
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide maleate and one or more associated
compounds having the following structures:

##STR00001## ##STR00002## ##STR00003##

[0020] The present invention also provides a pharmaceutical composition
for the inhibition of HER-2 kinase activity comprising a
therapeutically-effective amount of
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide maleate and a pharmaceutically
acceptable carrier. The pharmaceutical composition may also contain one
or more of the associated compounds discussed above. The maleate salt may
be in an anhydrous form, a monohydrate form, and combinations of these
forms.

[0021] The present invention also provides a method for preventing,
treating, or inhibiting cancer by administering a
therapeutically-effective amount of
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide maleate to a subject. The subject
may be a mammal, and more specifically, a human. The maleate salt may be
administered in its anhydrous form, monohydrate form, or partially
hydrated form. One or more of the associated compounds discussed above
may also be administered during this method.

BRIEF DESCRIPTION OF THE DRAWINGS

[0022]FIG. 1. The XRD scans of two crystalline forms of
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide maleate, anhydrous Form I and
monohydrate Form II.

[0028]FIG. 7. XRD scans of Form II before and after exposure to a
relative humidity of 50-60% at an ambient temperature of 20-25° C.
for 24 hours.

[0029]FIG. 8 XRD scans of Form I before and after exposure to relative
humidity of 50-60% at an ambient temperature of 20-25° C. for 24
hours.

DETAILED DESCRIPTION OF THE INVENTION

[0030] (E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy--
6-quinolinyl}-4-(dimethylamino)-2-butenamide is an irreversible inhibitor
to Her-2 (also known as ErbB-2 or neu) kinase, a member of the epidermal
growth factor receptor (EGFR) family. EGFR family members have been
implicated in tumorigenesis and associated with poor prognosis in tumor
types in humans. The structure of the
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide in the form of a free base is shown
below:

##STR00004##

[0031] The compound
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide in the form of a free base is
described in U.S. Pat. No. 6,288,082. The compound is classified, based
on the Biopharmaceutical Classification System, as a BCS Class IV
compound (low water solubility and low permeability). The free base has
low solubility in water, with a water solubility of about 1 μg/mL at
about pH 7. The water solubility increases with decreasing pH as the
compound becomes ionized. This compound is water soluble at
gastrointestinal pH, and dissolution is not rate limiting. There is a
need for a form of this compound with improved physicochemical
properties.

[0032] The present invention provides a water-soluble acid addition salt
form of (E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethox-
y-6-quinolinyl}-4-(dimethylamino)-2-butenamide. The free base compound is
capable of forming salts with a variety of pharmaceutically suitable
acids. Pharmaceutically suitable acids include, but are not limited to
for example, acetic, fumuric, maleic, methanesulfonic, succinic,
sulfuric, tartaric, and p-toluenesulfonic acid. The physicochemical
properties of each acid addition salt form were evaluated to screen for
an optimal pharmaceutical salt form, as shown in Table 1.

[0033] Of the nine salts, the maleate salt exhibited advantageous
physicochemical properties. The maleate salt was crystalline and less
hygroscopic. The mesylate salt was hygroscopic and less crystalline. The
tosylate salt was even less attractive, primarily due to its higher
molecular weight and safety concerns. Although the acetate "salt"
appeared to be crystalline. NMR revealed that the product prepared from
acetic acid was in fact not a salt. The fact that the product prepared
from acetic acid was insoluble in water with a resulting alkaline pH
confirmed that it largely retained the free base properties.

[0034] The (E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-eth-
oxy-6-quinolinyl}-4-(dimethylamino)-2-butenamide maleate salt is
crystalline and has higher solubility in water as compared to the free
base as shown in Table 2.

[0035] A comparison of the systemic exposure (SE) data for
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide has been conducted on data
extracted from multiple preclinical studies in the rat. The analysis of
these data indicated that, in the rat, administration of the compound as
the maleate salt provided a two-fold increase in AUC (area under
concentration), as compared to the free base, when administered at a dose
range of 5 to 45 mg/kg. The systemic availability of the compound as the
free base was relatively low (20%), and the presence of significant
amounts of drug in the feces could be attributed to poor absorption. The
increased solubility of the maleate salt appears to enhance the
absorption of the compound in the rat. Table 3 presents the plasma
compound mean AUC and Cmax data observed in rats.

[0037] In addition to exhibiting poor water solubility, the compound
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide in the form of a free base
interacts with emectic receptors in the stomach, giving rise to diarrhea
in mammals. The maleate salt of
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide, however, unexpectedly mitigates
such problems and minimizes emectic receptor interactions in mammals.

[0038] The maleate salt is prepared by mixing
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide (the free base) with maleic acid
and dissolving the mixture in a water-alcohol solution at an elevated
temperature. The resulting solution is cooled and the cooled solution
contains (E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-etho-
xy-6-quinolinyl}-4-(dimethylamino)-2-butenamide maleate. According to one
embodiment,
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide maleate is prepared by combining
maleic acid and the free base in a solution of water and n-propanol, as
described in Scheme 1.

##STR00005##

[0039] The reaction of the free base and maleic acid occurs at an elevated
temperature of from about 40° C. to about 60° C.,
preferably between about 40° C. to about 50° C. The ratio
of water:n-propanol may vary, for example between about 1:10 to about
1:5, and the optimal ratio of water:n-propanol is about 1:9. The
water-alcohol solution may comprise from about 5% to about 20% by volume
water and from about 80% to about 95% by volume alcohol. The alcohol may
be n-propanol. In one embodiment, the water-alcohol solution comprises
about 10% by volume water and about 90% by volume n-propanol. The volume
of the solvent solution may be between about 8 to about 25 volumes,
including about 10 to about 12 volumes. About 1.0-1.2 equivalents of
maleic acid is used per equivalent of the free base, preferably about
1.03 equivalents of maleic acid per equivalent of the free base.

[0040] The resulting solution of the maleate salt may be clarified by
filtration prior to cooling. The cooling step may be continued until the
solution reaches a temperature of about 45° C. or less, including
a temperature of about 39° C. or less, and more preferably to
about 30° C. or less. In one embodiment, the solution is filtered
after cooling to about room temperature, preferably from about 23°
C. to about 25° C. Typically, the maleate salt begins to
crystallize out of solution once the temperature reaches 37° C. or
below. The solution may be allowed to sit for at least 12 hours,
preferably about 12 to about 15 hours at room temperature, and is then
filtered and washed to recover the crystalline maleate salt product. The
resulting filter cake may be washed with the same or a different
water-alcohol solution to obtain the product. The product may be dried to
obtain crystalline
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide maleate. At this point, the maleate
salt product recovered and isolated is typically in the form of the
monohydrate form of the maleate salt.

[0041] The product may be dried under vacuum with heating to make the
anhydrous form of the maleate salt (Form I) at about 70 to about 95%
yield, preferably about 80 to about 95% yield. This product is usually
better than about 98% pure, and often about 99% pure. Typically, the
drying process is performed over about 12 to about 48 hours to get
complete conversion of the anhydrous form of the maleate salt to the
monohydrate form of the maleate salt (Form II).

[0042] Shorter drying times generally result in mixtures of the two
crystalline forms. The drying process is often performed at temperatures
greater than room temperature. In one embodiment, drying of the maleate
salt is performed at a temperature greater than about 30° C.,
preferably from about 40° C. to about 60° C., and in
another embodiment at about 50° C.

[0043] The maleate salt is soluble in many polar solvents, which will be
known to one skilled in the art, but dimethyl sulfoxide (DMSO) is often
used if a small solvent volume is desired. The DMSO solution can be
heated to about 45° C. to about 60° C. to further enhance
solubility. Once the anhydrous maleate salt is in solution, water may be
added, typically quickly, causing the crystallization that provides the
crystalline monohydrate form upon filtration. The anhydrous salt may be
dissolved in a solvent, for example DMSO, and to this solution may be
added an aqueous solution of water and an organic solvent, for example
such as tetrahydrofuran (THF), isopropanol (IPA), n-propanol, acetone,
ethanol, methanol, and acetonitrile. In one embodiment, the organic
solvent used is IPA, in another embodiment it is n-propanol, and in a
third embodiment a mixture of these two organic solvents is used. The
water content of the aqueous solution can be as little as 5%, but may be
about 7.5% or greater, and in one embodiment is between about 10% and
about 15%. The resulting solution then may be allowed to sit for up to
about 24 hours, and in one embodiment is allowed to sit for between about
12 hours and about 24 hours, to allow for crystallization to occur.
Filtration of the mixture yields a crystalline monohydrate form of the
maleate salt. For purposes of this invention, the term "organic solvent
and water" refers to a solution of an organic solvent, such as for
example tetrahydrofuran (THF). DMSO, methanol, ethanol, isopropyl alcohol
or acetonitrile, and water wherein the organic solvent comprises greater
then 50% of the solution by volume.

[0044] The invented maleate salt of
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide was isolated in three different
crystalline forms: an anhydrous form (Form I), a monohydrate form (Form
II) and a partially hydrated form (Form III), which comprises a mixture
of Form I and Form II.

[0045] According to one embodiment, the anhydrous form of
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide maleate (Form I) is obtained as a
crystalline solid by drying the reaction product of
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide and maleic acid. Drying includes
air drying, heating and drying under reduced pressure. In an alternative
embodiment, the anhydrous form of
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide maleate (Form I) is obtained as a
crystalline solid by drying the monohydrate form of
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide maleate (Form II).

[0046] The isolated crystalline form of anhydrous
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide maleate (Form I), is characterized
by differential scanning calorimetry (DSC), as exhibiting an onset
temperature in the range of about 196-204° C., at which melting
and decomposition occur.

[0048] According to one embodiment,
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide maleate is prepared in the form of
a crystalline monohydrate (Form II) by mixing anhydrous
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide maleate (Form I) with an organic
solvent and an amount of water and filtering crystalline monohydrate that
precipitates from the mixture.

[0049] In a separate embodiment,
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide maleate in the form of a
crystalline monohydrate (Form II) is prepared by mixing anhydrous
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide maleate (Form I) with an organic
solvent; adding a solution comprising an amount of water in an organic
solvent; and filtering crystalline monohydrate that precipitates from the
mixture.

[0050] In another embodiment,
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide maleate in the form of a
crystalline monohydrate (Form II) is prepared by mixing anhydrous
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide maleate (Form I) with an organic
solvent comprising an amount of water over a period of days and filtering
crystalline monohydrate that precipitates from the mixture. The period of
days is suitably about 1-20 days.

[0051] The isolated crystalline form of
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide maleate monohydrate (Form II),
exhibits water loss at about 50° C., as measured by DSC, and is
characterized by a water content of about 2.5 to 2.7% by weight, as
measured by thermal gravimetric analysis (TGA), based on the weight of
the compound as a monohydrate. The water content of the monohydrate form
of the maleate salt was also measured by Karl Fischer titration.

[0053] As used herein, the term isolated means that more than 50% of the
crystalline
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide maleate salt present is one of
Forms I and U. In one embodiment, at least 70% of the crystalline
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide maleate salt present is one of
Forms I and II. In a second embodiment, at least 80% of the maleate salt
present is one of Forms I and II. In a third embodiment, at least 90% of
the maleate salt present is one of Forms I and II.

[0054] The two crystalline forms of
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide maleate, exhibit distinct XRD
patterns and peaks. The XRD pattern for each maleate salt form is unique
to that salt form. The XRD patterns of Forms I and II were determined by
using techniques and equipment known to those skilled in the art of
analytical chemistry and X-ray crystallography. XRD patterns were
produced using powder samples and are comprised of a set of diffraction
peaks, which can be expressed in 2 theta angles, d-spacing and/or
relative peak intensities. The XRD patterns are shown in FIGS. 1, 5, 6,
7, and 8. Collection parameters for the X-ray data provided in FIGS. 1, 7
and 8 were as follows: voltage 40 kV; current 40.0 mA; 5.00-30.00 degree
scan range; Bruker D8 Advance instrument; scan step size 0.01°;
total scan time 30 minutes; using a Vantec-1 detector and Ni filter. The
X-ray data in FIGS. 5 and 6 were collected as follows: voltage 30 kV;
current 15 mA; 3-40 degree scan range; 2.00°/min; Rigaku Miniflex
bench top X-ray diffractometer.

[0055] The two-theta diffraction angles and the corresponding d-spacing
values account for the positions of the peaks found in a XRD pattern.
D-spacing values are calculated with observed two theta angles and copper
Kα1 wavelength using the Bragg equation. Variations in these
numbers can result from using different diffractometers and also from the
method of sample preparation. However, more variation can be expected for
the relative peak intensities. Therefore, identification of the various
forms should be based upon the observed two-theta angles and the
d-spacings, and less importance should be given to the intensities. One
skilled in the art would understand that the XRD patterns of Forms I and
II obtained as described herein could contain additional peaks.
Additionally, a skilled artisan would recognize that whether all the
peaks are observed for a given form may be highly dependent on the
concentration level of the form. FIG. 1 illustrates XRD scans of the two
crystalline forms of the maleate salt, Form I and II. The crystalline
anhydrous maleate salt form, Form I, is shown on the bottom, while the
crystalline monohydrate form of the maleate salt, Form II, is shown on
top.

[0056] The relative stability and hygroscopicity of the two crystalline
forms of the maleate salt was studied in detail by dynamic vapor sorption
(DVS). The anhydrous form of the maleate salt absorbs water easily and
converts to the crystalline monohydrate form of the maleate salt. Upon
drying or a drop in the relative humidity, the crystalline monohydrate
form of the maleate salt converts to the anhydrous form of the maleate
salt, as summarized in FIG. 2. FIG. 2 is a dynamic vapor sorption
isotherm plot which shows that
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide maleate, Form I, gains moisture
above 40% relative humidity (RH), especially at 60% RH and above. FIG. 2
also shows that Form II loses water at 20% RH and below, especially at
10% RH and below. DVS was performed under the following conditions: RH
was set at 0%, 30%, 52.5%, 75% and 90%, with the sample exposed for 3
hours at each RH for two full cycles.

[0057] The two crystalline forms of
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide maleate exhibit distinct DSC
traces. A DSC plot of both Form I and Form II of the maleate salt is
summarized in FIG. 3. Form I of the maleate salt exhibits one endothermic
peak, indicating a transition temperature of 202.49° C. Form II of
the maleate salt exhibits two endothermic peaks, a broad endotherm having
an onset temperature of 55° C. corresponding to loss of water and
a second endotherm indicating a transition temperature of 202.81°
C. The transition temperatures are observed in the range of about
196-204° C. at which melting and decomposition occurs. DSC data,
transition temperatures and heat flow, were collected using a TA
instrument model Q1000 with the following parameters: 50 mL/min purge gas
(N2); scan range 40 to 240° C., scan rate 10° C./min.
Pure, crystalline solids have a characteristic transition temperature,
the temperature at which point the substance changes state, in the
present case the solid transitions to a liquid. The transition between
the solid and the liquid is so sharp for small samples of a pure
substance that transition temperatures can be measured to 0.1° C.
Because it is difficult to heat solids to temperatures above their
transition temperatures, and because pure solids tend to transition over
a very small temperature range, transition temperatures are often used to
help identify compounds. Measurements of the transition temperature of a
solid can also provide information about the purity of the substance.
Pure, crystalline solids transition over a very narrow range of
temperatures, whereas mixtures transition over a broad temperature range.
Mixtures also tend to transition at temperatures below the transition
temperatures of the pure solids.

[0058] TGA data of the monohydrate and anhydrous forms of the maleate salt
are summarized in FIG. 4. Form II of the maleate salt is characterized by
a water content of about 2.5 to 2.7% by weight, as measured by TGA, based
on the weight of the compound as a monohydrate. TGA data were collected
using a TA Instrument Model Q. A heating rate of 10° C./min
between 30-220° C. was used and the TGA chamber was under 40
mL/min flow of nitrogen.

[0059] A third crystalline form of the maleate is salt is observed and
referred to as the partial hydrate (Form III), as observed from XRD. The
partial hydrate is a mixture of Form I and Form II of the maleate salt.
The partially hydrated
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide maleate (Form III), is
characterized by a water content of about 0.8 to about 2.4% by weight,
including about 1.5% to about 2.3% by weight, based on the weight of the
compound.

[0060]FIG. 5 includes an XRD scan of each of anhydrous Form I,
monohydrate Form II and partial hydrate Form III of
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide maleate after exposure of the
anhydrous form of the maleate salt to a relative humidity of 75% at an
ambient temperature of 20-25° C. for 22 days.

[0061]FIG. 6 is an XRD scan of two batches of crystalline
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide maleate in Form I. The anhydrous
form of the maleate salt absorbs water and partially converts to the
monohydrate form of the maleate salt at an ambient temperature of
20-25° C. over 24 hours. The monohydrate form of the maleate salt
is relatively stable at an ambient temperature of 20-25° C. for 24
hours. FIG. 7 illustrates an XRD scan of crystalline
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide maleate in Form II, before and
after exposure to relative humidity of 50-60% at an ambient temperature
of 20-25° C. for 24 hours. Exposing the monohydrate form of the
maleate salt to higher temperatures (>50° C.) or heating under
reduced pressure promotes water loss and full conversion back to the
anhydrous form of the maleate salt.

[0062] Form I, the anhydrous form, is readily converted to the monohydrate
form, Form II. Form I can absorb water and convert partially to the
monohydrate at a temperature of 20-25° C. and a relative humidity
(RH) of 50-60% over time, as shown in FIG. 8. FIG. 8 is an XRD scan of
crystalline
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide maleate in Form I before (lower
scan), and after (upper scan) exposure to relative humidity of 50-60% at
room temperature of 20-25° C. for 24 hours. Hydrate peaks appear
in the upper scan, indicating that the crystals absorb water under these
conditions.

[0063] The stability of both forms of the maleate salt of
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide was evaluated in closed and open
containers at 40° C. and 75% RH. Both Form I and Form II remained
stable for 6 months under these conditions. In the open containers, the
anhydrous form of the maleate salt rapidly absorbed one mole of water to
form the monohydrate form of the maleate salt. Samples in the closed
containers remained dry. HPLC purity analysis indicated no significant
increase in degradation products in both open and closed conditions for
up to 6 months. The data is summarized in Table 5.

[0064] Reactive crystallization of the free base with maleic acid in
different solvents was performed to determine which crystalline form(s)
of the maleate salt resulted. Table 6 illustrates the results of the
crystallization process in a mixture of n-propanol and water at various
operating conditions. The wet cake in all experiments contains the
monohydrate form of the maleate salt, which converts to the anhydrous
form of the maleate salt after drying.

[0066] One solvent that appreciably dissolves the
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide maleate salt is dimethylsulfoxide
(DMSO). Cooling, anti-solvent and evaporative crystallization were
performed in mixtures of DMSO and isopropanol ort-butyl methyl ether
(tBME). The approach led to the decomposition of the solute in many
cases. Anti-solvent and evaporative crystallization did not result in any
new crystalline forms, as summarized in Tables 8 and 9.

[0067] According to one embodiment, one way to convert anhydrous Form I
into monohydrate Form II is by dissolving the salt into a solution of an
organic solvent, for example such as THF, isopropanol (IPA), n-propanol,
acetone, ethanol, methanol, and acetonitrile, and water, where in the
water present is about 5% to about 20% by volume, though typically the
water present is about 10% to about 15% by volume. This solution may be
heated to increase solubility of the maleate salt; in one embodiment it
is heated to about 45° C. or greater, in another embodiment it is
heated to about 60° C. The solution is then allowed to sit for a
period of hours to allow for crystallization, and the crystals are then
filtered to give monohydrate Form II (see Table 6). In one embodiment the
solution is allowed to sit for between about 12 and about 24 hours before
filtration.

[0068] According to a separate embodiment, Form I is converted to Form II
by re-slurrying it in organic solvent containing water and allowing the
solution to stand exposed to the room temperature for several days, as
shown in stability studies summarized in Table 10. This conversion will
take place even in anhydrous solvents that have absorbed up to 1% water
because anhydrous Form I readily absorbs moisture, as evidenced by FIG.
8. In one embodiment the re-slurry is allowed to stand for about 14 days.

[0069] The present invention is also directed to compounds associated with
the free base or the maleate salt of
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide, or the methods of this invention.
One or more of these associated compounds may be found in the cooled
solution in a process of this invention. Since these compounds may not be
separated from the maleate salt, a pharmaceutical formulation prepared
with the maleate salt may contain one or more of these compounds.

[0070] Formulations of the maleate salt were prepared and stored in
40° C./75% RH stability chambers for six months and in a
56° C. oven for one month. Samples were periodically pulled for
testing. Samples were dissolved in 50/50 volume/volume acetonitrile/water
with a concentration at about 0.5 mg/mL. The solutions were assayed
directly using LC/MS methodology to identify any degradation products and
impurities (referred to herein as associated compounds) at six-months.
Structures of the associated compounds, detected by LC/MS are listed in
Table 11. Notably, the amount of the degradation product associated with
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide maleate is reduced by the
production method of the present invention.

[0083] Crystalline forms of the maleate salts of the present invention are
useful for preventing, treating, or inhibiting inflammation or cancer by
administering a therapeutically-effective amount of
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide maleate to a subject. The subject
may be a mammal, and more specifically, a human. The maleate salt may be
administered in its anhydrous form, monohydrate form or partially
hydrated form. One or more of the associated compounds discussed above
may also be administered during this method.

[0084] Crystalline forms of the maleate salts of the present invention are
useful for preparing pharmaceutical compositions for the inhibition of
HER-2 kinase activity, which is linked to the treatment of cancer. The
formulations contain a therapeutically effective amount of
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide maleate and a pharmaceutically
acceptable carrier. The pharmaceutical composition may be administered in
its anhydrous form, monohydrate form or partially hydrated form. One or
more of the associated compounds discussed above may also be administered
during this method.

[0085] Pharmaceutical compositions and formulations of the present
invention may be useful in the treatment of one or more of breast cancer,
ovarian cancer, epidermoid tumors, colon cancer, prostate cancer, kidney
cancer, bladder cancer, larynx cancer, esophagus cancer, stomach cancer,
and lung cancer. According to one embodiment, the maleate salt is
particularly useful in the treatment of breast cancer and/or ovarian
cancer.

[0086] The pharmaceutical compositions and formulations including maleate
salt forms of the invention may be administered orally, by intralesional,
intraperitoneal, intramuscular or intravenous injection; infusion;
liposome-mediated delivery; topical, nasal, anal, vaginal, sublingual,
uretheral, transdermal, intrathecal, ocular or otic delivery. One mode of
administration for the compound of the invention is the unit dose form.
Suitable unit dose forms include tablets, capsules and powders in sachets
or vials. The crystalline compounds of the present invention can be
administered orally. Such compounds may be administered from 1 to 6 times
a day, more usually from 1 to 4 times a day. The effective amount will be
known to one of skill in the art; it may also be dependent upon the form
of the compound, the mode of administration and the serverity of the
condition being treated. One of skill in the art could routinely perform
empirical activity tests to determine the bioactivity of the compound in
bioassays and thus determine what dosage to administer. However, in
general, satisfactory results can be obtained with compounds of the
present invention when dosed daily in the range of about 0.5 mg/kg to
about 1000 mg/kg of body weight, but usually the effective dosage amount
is between about 1 mg/kg to about 300 mg/kg per day.

[0087] The crystalline forms of maleate salts of the invention may be
formulated with conventional excipients, such as fillers, disintegrating
agents, binders, lubricants, flavoring agents, color additives, and
carriers. The carrier may be a diluent, an aerosol, a topical carrier, an
aqueous solution, a nonaqueous solution or a solid. The carrier may be a
polymer or a toothpaste. A carrier in this invention encompasses any of
the standard pharmaceutically accepted carriers, such as phosphate
buffered saline solution, acetate buffered saline solution, water,
emulsions such as an oil/water emulsion or a triglyceride emulsion,
various types of wetting agents, tablets, coated tablets and capsules.

[0088] If administered orally or topically, the crystalline forms of
maleate salts of the invention may be provided to a subject in different
carriers. Typically, such carriers contain excipients such as starch,
milk, sugar, certain types of clay, gelatin, stearic acid, talc,
vegetable fats or oils, gums, or glycols. Specific carriers are typically
selected based upon the desired method of delivery, for example,
phosphate buffered saline (PBS) could be used for intravenous or systemic
delivery and vegetable fats, creams, salves, ointments or gels may be
used for topical delivery.

[0089] The crystalline forms of maleate salts of the present invention may
be delivered together with suitable diluents, preservatives,
solubilizers, emulsifiers, adjuvants and/or carriers useful in treatment,
inhibition or prevention of neoplasm. Such compositions are liquids or
lyophilized or otherwise dried formulations and include diluents of
various buffer content (for example, Iris-HCl, acetate, phosphate), pH
and ionic strength, additives such as albumins or gelatin to prevent
absorption to surfaces, detergents (for example, TWEEN® 20, TWEEN®
80, PLURONIC® F68, bile acid salts), solubilizing agents (for example,
glycerol, polyethylene glycerol), anti-oxidants (for example ascorbic
acid, sodium metabisulfate), preservatives (for example, thimerosal,
benzyl alcohol, parabens), bulking substances or tonicity modifiers (for
example, lactose, mannitol), covalent attachment of polymers such as
polyethylene glycol, complexation with metal ions, or incorporation of
the compound into or onto particulate preparations of hydrogels or
liposomes, micro-emulsions, micelles, unilamellar or multilamellar
vesicles, erythrocyte ghosts, or spheroblasts. Such compositions will
influence the physical state, solubility, stability, rate of in vivo
release, and rate of in-vivo clearance of the compound or composition.
The choice of compositions will depend on the physical and chemical
properties of the compound.

[0090] The crystalline forms of maleate salts of the invention also may be
delivered locally via a capsule that allows a sustained release of the
compound over a period of time. Controlled or sustained release
compositions include formulations in lipophilic depots (for example,
fatty acids, waxes, oils).

[0091] The crystalline forms of maleate salts of the invention can also be
dosed with other active compounds that would be of benefit to a patient
suffering from cancer, for example, other chemo agents or anti-biotics,
or in conjunction with radiation therapy. These active compounds can be
dosed with the compounds of the present invention simultaneously or in
sequence. The compounds of the present invention can also be formulated
to include the other active compound in the same dosage unit, for example
both could be contained within one pill, table or capsule. Some of the
possible types of active compounds that the compounds of the present
invention could be used in combination with are mitotic inhibitors, such
as taxol and vinblastine, alylating agents, such as cisplatin and
cyclophosamide, antimetabolites, such as 5-fluorouracil and hydroxyurea.
DNA intercalators, such as adriamycin and bleomycin, topoisomerase
inhibitors, such as etoposide and camptothecin, antiangiogenic agents,
such as angiostatin, and antiestrogens, such as tamoxifen.

[0092] This invention will be more fully described in conjunction with the
following specific examples, which should not to be construed as limiting
the scope of this invention. A skilled artisan will be able to
re-arrange, combine, modify, or eliminate steps in the exemplified
process, depending on process parameters and equipment.

[0093] Crude
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide free base (0.100 kg, 0.159 mole) is
rinsed with a 10% solution of USP purified water in n-propanol (0.082 kg,
0.10 L) followed by the addition of water:n-propanol solution (0.74 kg,
0.90 L). Maleic acid is added (0.0191 kg, 0.164 mole) and the mixture is
rinsed with 10% water:n-propanol (0.082 kg, 0.10 L). The mixture is
quickly heated to 50-60° C. and held for a minimum of 15 min,
until a solution is obtained. The hot solution is clarified through a
pre-heated 50-60° C., 0.2 Mm filter cartridge and the filtrates
are collected in a preheated 45-55° C., 2 L multi-neck flask. The
filter cartridge is rinsed through with 10% water:n-propanol pre-heated
to 45-55° C. (0.082 kg, 0.10 L). The solution is cooled over at
least one hour to 40° C. and held at that temperature for 12 hours
then cooled to room temperature (25° C.) over a minimum of four
hours and held at that temperature for at least two hours. The mixture id
filtered on a 12.5 cm diameter Buchner funnel for 5 min., then rinsed and
washed with pre-filtered 10% water:n-propanol solution (2×0.12 kg,
2×0.15 L). The cake is dammed and suction maintained until dripping
essentially stops, about 1 h.

Example 2

Preparation of
(E)-N-{4-[3-chloro-4-(2-pyridinylmethoxy)anilino]-3-cyano-7-ethoxy-6-quin-
olinyl}-4-(dimethylamino)-2-butenamide maleate, Form I